Snu620

Five gastric cancer cell lines (SNU5, SNU620, SNU638, Hs746T, and MKN45) were obtained from the Korean Cell Line Bank. Normal immortalized cell lines (HFE145) were obtained from Dr. Ashktorab (Howard University, Washington, DC, USA). Two non-small cell lung cancer (NSCLC) cell lines, EBC-1 and H1993, were purchased from the Japanese Research Resources Bank and the American Type Culture Collection, respectively. All cell lines were authenticated by short tandem repeat analyses in Korea Genome Information Institute and were checked for mycoplasma using the e-Myco VALID Mycoplasma PCR Detection Kit (#25245; iNtRon Biotechnology, Inc., Seongnam-si, Korea).

Twelve gastric cancer cell lines (six were established from male patients: NCI-N87, SNU-484, SNU-1, SNU-638, KATOⅢ, MKN-1; six were established from female patients: SNU-5, MKN-28, SNU-216, SNU-16, AGS, SNU-620) were obtained from Korean Cell Line Bank (KCLB; Seoul, Korea). All of the cells were cultured in RPMI 1640 (Gibco BRL, Grand Island, NY, USA) supplemented with 10% fetal bovine serum (FBS) and antibiotics (100 U/mL penicillin and 100 g/mL streptomycin; Gibco BRL).

Four human GC cell lines (NUGC-2, SNU-16, SNU-620, and SNU638), obtained from the Korean Cell Line Bank (Seoul, South Korea) were grown in RPMI-1640 medium supplemented with 10% fetal bovine serum and 1% antibiotic solution containing penicillin and streptomycin. Cells were incubated at 37ºC in a humidified atmosphere of 5% CO₂. Genomic DNA was extracted from these cell lines using a QIAamp DNA Mini Kit (QIAGEN).

To profile the methylome and miRNome of GC, 3 GC and adjacent normal tissue samples were obtained, along with informed consent, from Pusan National University Yangsan Hospital in Korea. EpCAM+/CD44+ GC cells were isolated from the 3 GC tissues according to a previously described protocol.10 To measure the protein expression levels of miR‐1271 target genes in GC tissues, 3 paired normal tissues and GC tissues were obtained, along with informed consent, from Chungnam National University Hospital in Korea. These studies were approved by the Internal Review Board at the corresponding hospitals, and all the experiments were performed in accordance with the relevant guidelines and regulations.
Nine GC cell lines (SNU‐1, SNU‐216, SNU‐484, SNU‐601, SNU‐620, AGS, KATO III, MKN1, and MKN74) and 293T cells were purchased from Korean Cell Line Bank (Seoul, Korea). The GC cell lines and 293T cells were cultured in Roswell Park Memorial Institute (RPMI) 1640 medium and Dulbecco’s Modified Eagle’s Medium (DMEM, WELGENE, Daegu, Korea), respectively, supplemented with 10% fetal bovine serum (FBS) (HyClone, Logan, UT, USA) in a CO₂ incubator, and the cell lines were authenticated by short tandem repeat (STR) DNA profile analysis performed by the Korean Cell Line Bank facility.

Gastric cancer cell lines (SNU1, SNU16, SNU216, SNU620, SNU638, and N87) were purchased from the Korean Cell Line Bank (Seoul). Cells were cultured in RPMI1640 supplemented with 25 mM HEPES, 10% fetal bovine serum (FBS), and 100 μg/ml of penicillin/streptomycin (1 × P/S) in 5%CO₂/95% air at 37°C. Cells were transfected with siRNA using DharmaFECT reagent 1 or 3 (Dharmacon, Lafayette, CO), according to the manufacturer’s instructions. The sequences of siRNA used were as follows: MED30 siRNA (Bioneer, Daejeon, Korea), 5’-CGA GCA ACU UAU UCC AUA U(dTdT)-3’, 5’-GCU GCC AAA UGG UGU CAC U(dTdT)-3’, and 5’-CGA GAA AUU GCU GAA GUA A(dTdT)-3’; scrambled (SCR) siRNA (Dharmacon, Lafayette, CO), 5’- GAU CCG CAA AAG AGC GAA A(dTdT)-3’.